DNA SEQUENCING OF UV INDUCED CHROMOSOMAL MUTATIONS

紫外线诱导的染色体突变的 DNA 测序

• 批准号: 3282087
• 负责人: Christopher W Lawrence
• 金额: $ 7.93万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-12-01 至 1986-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3282087
• 关键词: Escherichia coli; bacterial DNA; bacterial genetics; chromosome aberrations; chromosome deletion; genetic manipulation; genetic mapping; lac operon; nucleic acid sequence; plasmids; radiation genetics; ultraviolet radiation

The long term objective of this project is to describe the molecular processes responsible for the induction of mutations when cells are treated with DNA-damaging mutagens. The immediate aims are to characterize the population of nucleotide sequence changes induced by ultraviolet light and related mutagens in chromosomal DNA of E. coli, to separately distinguish among them those that are targeted from those that are untargeted, and to determine the role of certain key gene functions in the production of the various types of mutations. This will be achieved by isolating mutations from lacI+ to lacI- for genes on an F' pro lac transferred to an independently treated Deltapro lac recipient, mapping these mutations to one of five large regions using deletions, transferring the mutations by recombination to an f1 lacI+ lacZ'Delta47 hybrid phage and sequencing them by means of the Sanger method. This will provide information about the capabilities of the altered replication complex, thought to exist in recA induced cells, when confronted with template defects of various kinds or normal DNA, and on the base-pairing capacity, if any, of the lesions. Understanding the mode of action of these mutagens is relevant to health concerns of two kinds. Environmental agents of this type may represent a health hazard, both as mutagens and carcinogens, but the magnitude of this risk can be fully assessed only when their mechanism of action is described in detail. Further, certain congenital diseases appear to be the consequence of DNA repair deficiencies, but these are poorly understood.

该项目的长期目标是描述分子 处理细胞时负责诱导突变的过程 破坏DNA的诱变剂 直接的目的是描述 紫外光诱导的核苷酸序列变化群体， E.大肠杆菌，分别区分 他们中的许多人，都是有目的的， 确定某些关键基因功能在生产中的作用， 各种类型的突变。 这将通过分离突变来实现 从lacI+到lacI-，用于转移到 独立治疗的Deltapro lac受体，将这些突变映射到 使用缺失的五个大区域之一，通过 重组到f1 lacI+ lacZ ′ Delta 47杂合噬菌体上并测序 通过桑格方法。 这将提供有关 被认为存在于recA中的改变的复制复合体的能力 诱导细胞，当面对各种模板缺陷或 正常的DNA，和碱基配对能力，如果有的话，病变。 了解这些诱变剂的作用方式与健康有关 有两种担忧。 这种类型的环境因素可能代表 健康危害，既作为诱变剂和致癌物，但这一规模 只有在描述其作用机制时，才能充分评估风险 详细 此外，某些先天性疾病似乎是 这是DNA修复缺陷的结果，但人们对此知之甚少。